Original ResearchFull Report: Basic and Translational—LiverOncogenic Mutations in Armadillo Repeats 5 and 6 of β-Catenin Reduce Binding to APC, Increasing Signaling and Transcription of Target Genes
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Section snippets
Catalogue of Somatic Mutations in Cancer Database Analysis
To obtain the cancer-related CTNNB1 mutations depicted in Figure 1, we analyzed the COSMIC Web site (https://cancer.sanger.ac.uk/cosmic/gene/analysis?ln=CTNNB1), filtering it on amino acids 310–440. Data were updated until January 2019.
Plasmids and Construction
N-terminal FLAG-tagged β-catenin variants were constructed by using the pcDNA 5′ UT-FLAG vector, as previously described.13 Briefly, the constructs of WT (WT), S33Y, and exon 3 deletion were generated by using the Gibson assembly method New England Biolabs (NEB).
K335I and N387K β-Catenin Variants Are Potent Inducers of Hepatocellular Carcinoma Formation in Mice
To directly demonstrate the oncogenic potential of the β-catenin K335I and N387K variants, we hydrodynamically injected them in the tail vein of mice.21 WT β-catenin was taken along as control. All variants were cotransfected with c-Met because previous work showed that oncogenic β-catenin alone is insufficient for hepatocellular carcinoma (HCC) development.16,17 In strong contrast to the mice injected with WT β-catenin, all K335I/N387K injected mice became moribund 6–7 weeks after injection,
Discussion
The β-catenin signaling pathway is one of the most commonly deregulated pathways among cancers.1 In colorectal cancers, this is predominantly accomplished by inactivating APC mutations, whereas in liver cancers, for example, aberrant activation has been mainly attributed to activating mutations in the CTNNB1 gene (20%–25%).4,7,33, 34, 35 The exon 3–related β-catenin mutations acquire enhanced signaling activities by interfering with proper N-terminal phosphorylation and subsequent proteolytic
Acknowledgments
Author contributions: Pengyu Liu performed the majority of experimental work and data analysis and authored the manuscript; Binyong Liang, Manning Qian, and Xin Chen performed and coordinated the animal experiments involving hydrodynamic transfection; Menggang Liu, Marla Lavrijsen, and Shan Li assisted with the experiments; Joyce H.G. Lebbink performed all the structural protein analyses and authored parts of the manuscript; Maikel P. Peppelenbosch supervised the project and improved the
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Conflicts of interest The authors disclose no conflicts.
Funding This research was financially supported by a China Scholarship Council PhD fellowship to Pengyu Liu (file no. 2014 0822 0029), Shan Li (file no. 2014 0806 0053) and Manning Qian (file no. 2018 0832 0464) and a National Institutes of Health grant (R01CA204586) to Xin Chen. Joyce Lebbink is supported by the gravitation program (024.001.028) from The Netherlands Organization for Scientific Research (NWO).
Author names in bold designate shared co-first authorship.